PCR技术应用于临床结核病的诊断研究

Detection of M.Tuberculosis PCR and Its Application in Diagnosis

采用PCR(聚合酶链反应)技术,选择一对寡核苷酸引物,扩增的靶基因为克隆的PH_重组标准人型结核分枝杆菌2.4kb DNA。其基因片段是结核杆菌所特有,单一拷贝基因,与其他非典型分枝杆菌的基因无同源序列。研究结果表明:①扩增了人型和牛型结核杆菌标准株基因DNA 158bp 片段,而其他14种非典型分枝杆菌和金黄色葡萄球菌、脓杆菌 DNA 未扩增出相应产物.②10倍系列稀释人型结核菌 DNA,检测敏感性相当于10～50菌数/ml痰样。③60例临床肺结核病人痰样进行 PCR 技术检测和常规生物法测定比较,前者阳性率为58.3%,后者检测率11.70%。而20例非结核痰样用两种方法检测全为阴性。提示:PCR 技术诊断结核病快速、简易、特异、敏感、高效,是在基因水平上检测的一种新技术。

This article introduces polymerase chain reaction(PCR)amplification DNA sequence specific of M.Tuberculosis for its application in diagnosis.We have been describe Mycobacterial and non-my- cobacterial reference bacterial strains and 60 cases clinical isolates of M.Tuberculosis and 20 cases non- Tuberculosis.the results suggested that:①there's a 1 58-bp DNA fragment of M bovis,and M.human by PCR.②DNA from less than 10 Mycobaterial cells can be detected using the PCR.③60 cases clinical isolates of M.Tuberculosis positive rate separately were 58.3% by PCR and 11.7% by cul- turing.The data reported here suggest that DNA amplification is a superior method with a high degree of sensitivity and specifity for the detection of Mycobacterial DNA sequences,the method of M.Tu- berculosis by PCR may be a very usefal and valuable tool for rapid detection of Mycobacterial in uncul- tured clinical specimens.